Objective To study the effect of acute non-isovolemic hemodilution (ANIH) plus tranexamic acid on bleeding and coagulation factors. Methods Forty-two patients with brain tumor under general anesthesia were randomly divided into group N and group T with 21 patients in each group. Group N was given ANIH , while group T was given tranexamic acid and ANIH. Bleeding, transfusion, urine volume were recorded. Coagulation factors and D-dimer were detected one day before and after surgery. Hemoglobin was recorded before and after ANIH and after auto-blood was transfused. Results There was less bleeding in group T. Hemoglobin in group T was higher after transfusion. No significant difference was found in Group T and group N in terms of urine volume and transfusion rate. Both the two groups had no difference on variation of coagulation factors. Conclusion ANIH with tranexamic acid has no significant effect on coagulation but produces synergetic effect on decreasing bleeding. They can be applied in surgery of brain tumor safely.

Purpose To compare the CTPA image quality by using contrast agent with different concentration at different injection rate so as to provide suitable contrast agent injection for patients.Materials and Methods A total of 346 patients with suspected acute pulmonary embolism who required to undergo CTPA examination were randomly assigned to high (370 mgI/ml) and low (320 mgI/ml) concentration groups,and each group was further divided into six subgroups with different velocity (3.0,3.2,3.4,3.6,3.8 and 4.0 ml/s).The CT value of the main pulmonary artery,right pulmonary upper lobe artery and right lung under leaf posterior basal segmental artery was measured.Results In the high concentration group,there were no significant differences in pulmonary artery average CT value,noise,single to noise ratio (SNR) and contrast to noise ratio (CNR) among the subgroups with different velocity (P＞0.05).In the low concentration group,the difference was not statistically significant in pulmonary artery average CT value (P＞0.05) among the subgroups with different velocity;however,the noise,SNR and CNR of 3.0 ml/s subgroup had significant differences compared with other subgroups (P＜0.05).There was no significant difference in average CT value of pulmonary artery between the subgroups with the same velocity in the two concentration groups (P＞0.05).In addition,except that the noise,SNR and CNR of 3.0 ml/s subgroup showed significant differences with other subgroups either in high concentration group or in low concentration group (P＜0.05),there were no significant differences in the above-mentioned parameters among other subgroups with the same velocity in both groups (P＞0.05).Conclusion Compared with high concentration contrast agent,the image obtained by using low concentration contrast agent shows no difference in pulmonary artery average CT value but with low iodine flow and iodine flow rate,which can reduce the risks of contrast media induced nephropathy (CIN) and contrast agent extravasation.

Objective To investigate the effect of acute non-isovolemic hemodilution in combination with tranexamic acid on cycle function blood gas and electrolytes with brain tumor surgery. Methods Forty-two patients undergoing brain tumor were randomly divided into two groups. Patients in group A received ANIH plus tranexamic acid , while patients in group B received ANIH alone. Collected blood was transfused before the end of surgery. HR、CVP、MAP,hemoglubin, blood gas and plasma electrolytes were respectively recorded before ANIH(T1), at 0 min (T1) and 1 h (T2) after ANIH, and at the end of operation (T4). Results There were no significant changes in HR, CVP, MAP. At T2, T3, T4, Hb, Hct in both two groups lower than those at T1(P <0.05); at T4, Hb, Hct in group A were higher than those in group B. There were no significant changes in pH , PaO2, PaCO2, BE between the both two groups. There were no significant changes in Na +, Cl-, Ca2+and K+between the both two groups. Conclusion ANIH has little effect on the cycle function and blood gas electrolyte. ANIH in combination with TA has a section blood effect. It can be used in the brain tumor operation with TA security.

Objective To investigate the effects of controlled hypotension (CH) combined with tranexamic acid (TA) on peri-operative blood loss and coagulation function in patients undergoing brain tumor surgery. Methods Forty patients undergoing brain tumor surgery were randomly allocated into group A and group B with 20 patients in each group. Patients in group A received CH alone, while patients in group B received CH combined with TA. Coagulation factors and d-dimer levels were measured 24 hours before and after surgery. Amount of blood loss, intravenous fluid transfused, urine output and postoperative drainage were recorded. Results D-dimer levels of 24 hours after surgery increased compared with that of 24 hours before surgery. In group B, the d-dimer level increased more than that of group A (P < 0.05). No significant difference was found in coagulation factor levels between group A and group B. Amount of blood loss, intravenous fluid transfused and postoperative drainage flows of patients in group B were lower than that in group A (P < 0.05). There were no significant changes in urine output and fluid infusion volume between two groups. Conclusion CH compared with TA can reduce perioperative blood loss in patients undergoing brain tumor surgery , with no obvious coagulant function abnormality. Collectively, it should be a safe and reliable method for clinical application.

Objective To determine the optimum harvest time of Scutellaria barbata D.Don with flavonoids contents as the index. Methods The content of total flavonoids in Scutellaria barbata D. Don was determined by ultraviolet-visible spectrophotometry with scutellarin as standard control, and scutellarin and cosmosiin in Scutellaria barbata D. Don were determined by HPLC. Results The contents of total flavonoids and scutellarin in Scutellaria barbata D.Don of different harvest time were complied with the quality standards in the China Pharma Copoeia, and reached the peakin blossom . The contents of total flavonoids, scutellarin and cosmosiin were (45.74±0.95) ,(5.58±0.16 ) and (17.39±0.42) mg?g-1 , respectively. Conclusion The Scutellaria Barbata D.Don may be collected at the flowering time with luxuriant foliage.

Objective:To evaluate the clinical efficacy of multi-disciplinary single center's CCCG-HB-2016 regimen in the treatment of hepatoblastoma (HB) in children.Methods:Clinical data of 36 HB patients treated with CCCG-HB-2016 program from Aug 2016 to March 2020 were analyzed.Results:These 36 patients included 20 boys and 16 girls. The serum AFP was all higher than 2 792 ng/ml,there was a correlation between AFP and tumor risk stratification ( H=14.973, P<0.05). Twenty eight cases (77.78%) were epithelial type and 8 cases (22.22%) were mixed epithelial mesenchymal type.All children were treated by tumor resection combined with chemotherapy, and there was a correlation between tumor risk stratification and surgical resection of liver lobe ( H=8.847, P<0.05). The probability of bone marrow suppression in the low-risk group was 58.33% (35/60),that in the intermediate-risk group was 73.49% (61/83) and in the high-risk group was 80.23% (69/86).All 36 cases were followed up to March 31, 2020,with an average follow-up of 21.9 months and the median survival was 22.5 months.The overall survival rate (OS) and event-free survival rate (EFS) were 97.2% and 83.3% respectively. Conclusions:The multidisciplinary CCCG-HB-2016 regimen was with a high success rate and along with a high incidence of bone marrow suppression.

Objective : To construct myeloid specific Spi1 gene knockout mice and analyze their genotypes , so as to provide animal model basis for the study of pathological mechanism of diseases and drug targets . Methods : According to the principle of CRISPR/Cas9 technology and C re/LoxP system , sgRNA and Donor vectors were de signed and constructed . The transcript of Exon 2 ( Exon 2) was used as the knockout region , and Loxp elements were placed on both sides of Exon 2 . Cas9 protein , sgRNA and Donor vector were mixed and microinj ected into the fertilized eggs of C57BL/6J mice , the fertilized eggs were transplanted into the uterus of C57BL/6J pregnant female mice , and F0 generation was obtained after 19 ~ 20 days . Positive F0 mice were mated with C57BL/6J mice to ob tain stable F1 Spi1 flox/ + mice . Spi1 flox/ + mice of F1 generation were selfed to obtain Spi1 flox/flox mice . Spi1 flox/flox mated with Lyz2-Cre + mice to obtain Spi1 flox/ + /Lyz2-Cre + mice , and then mated with Spi1 flox/flox , the Spi1 flox/flox/Lyz2-Cre + mice were myeloid specific Spi1 gene knockout ( KO) mice . Spi1 flox/flox/Lyz2-cre - mice were used as wild type (WT) mice . DNA of WT and KO mice was extracted , and the genotypes were identified by agarose gel electro phoresis after PCR amplification . Western blot was used to detect the expression of spleen focus forming virus proviral integration oncogene , Spi - 1 /purine rich box - 1(PU . 1) in immune cells of WT and KO mice . Results: The results of PCR identification showed that the genotype of mice with only 220 bp amplified by flox primer was Spi1 flox/flox homozygote , and the genotype of mice with 700 bp amplified by Lyz2-Cre primer was Lyz2-Cre + . Western blot showed that compared with WT group , the protein PU . 1 was not expressed in bone marrow derived macropha ges (BMDMs ) and peritoneal macrophages (PM) in KO group (P < 0.01) . There was no significant difference of statistics in the expression level of PU . 1 in T cells between KO mice and WT mice . The results of PCR and West ern blot showed that myeloid specific Spi1 KO mice were successfully constructed . Conclusion The myeloid spe cific Spi1 gene KO mice are successfully constructed and identified , which provides animal model basis for further revealing the potential mechanism of PU . 1 inimmune regulation .

Objective : To construct Csf1r-CreERT2 R26REYFP reporter gene mice and assess the efficacy of Csf1r- CreERT2 -mediated enhancement of CSF1R in CD45 + cells labeled with yellow fluorescein protein EYFP． Methods: Csf1r-CreERT2 mice were crossbred with R26REYFP homozygous mice，and Csf1r-CreERT2 R26REYFP mice were identified through PCR and Western Blot analyses．Flow cytometry was employed to evaluate CSF1R tag-efficiency in CD45 + cells across different mouse tissues following tamoxifen induction. Results : Csf1r-CreERT2 R26REYFP reporter gene mice were acquired．In addition，it was found that Csf1r-CreERT2 -mediated EYFP could effectively mark CSF1R in various tissues of mice and CD45 + cells in different locations．Compared to the R26REYFP group，the highest labeling efficiency was observed in the brain tissue (P＜0. 001) ，the lowest in the thymus tissue (P＜0. 05) ，and no sig- nificant difference was observed in the spleen tissue． Conclusion Adult Csf1r-CreERT2 mice and R26REYFP mice are effective ways to obtain Csf1r-CreERT2 R26REYFP induced conditional fluorescence mice．Csf1r-CreERT2 can mediate EYFP to effectively trace CSF1R in CD45 + cells in different parts of mice.

Objective : To breed and identify the T lymphocyte-conditional Spi1 knockout mice for the further in- vestgation of the specific role of Spi1-encoded protein PU. 1 . Methods : The Lck-Cre mice were mated with Spi1 flox/flox mice to obtain Lck-Cre ×Spi1 flox/flox mice (T lymphocyte-specific Spi1 knockout mice) , and the genotype was determined by polymerase chain reaction (PCR) and agarose gel electrophoresis . Magnetic beads were used to sort out the splenic T lymphocytes , and the knockdown efficiency of PU. 1 in T cells was detected by Western blot , quantitative real-time PCR ( qPCR) and flow cytometry. Results : The Lck-Cre ×Spi1 flox/flox mouse genotype was stably inherited . Compared with Spi1 flox/flox mice , the expression level of PU. 1 was significantly reduced in splenic T cells of Lck-Cre ×Spi1 flox/flox mice . Conclusion In this study , the T lymphocyte-specific Spi1 knockout mice was successfully constructed by applying Cre/LoxP system and CRISPR/Cas9 technology , which provided a reliable an- imal model for the subsequent experiments of the specific role of PU. 1 in T cell-related diseases .

Objective: To obtain chimeric antigen receptor macrophages ( CAR-M) targeting HER2 stably transfected． Methods : CAR lentivirus vector targeting HER2 was constructed and infected with human monocytic leukemia cell line (THP-1) ．CAR THP-1 cells with green fluorescent labeling were selected by sorting flow cytometry and continued to be cultured in vitro．The CAR THP-1 cells targeting HER2 were co-cultured with the endometrial cancer cell line Ishikawa with negative and positive HER2 expression，and their targeted phagocytosis of CAR-M to HER2 positive tumor cells was detected by imaging flow cytometry ，and the targeted phagocytosis efficiency of CAR-M to HER2 positive tumor cells was detected by flow cytometry. Results : CAR lentivirus infection with THP- 1 cells was less efficient ; After co-culture with cancer cells，flow cytometry and imaging flow cytometry showed that CAR THP-1 cells had enhanced phagocytosis of HER2 positive Ishikawa cells compared with the empty body group (P＜0. 01) ． Conclusion In this experiment，CAR THP-1 cell line targeting HER2 was established by constructing CAR lentivirus vector and transfecting THP-1 cells ，and it was proved that CAR THP-1 could phagocytize HER2 positive Ishikawa cells through specific targeting.